Composition comprising banyan tree, lotus, and clover serum fractions (hyperpigmentation)

ABSTRACT

The present invention relates to compositions comprising banyan tree, lotus, and clover serum fractions. A method of improving the appearance of a hyperpigmented spot may comprise the step of applying a composition comprising an effective amount of banyan tree serum fraction, lotus serum fraction, and clover serum fraction to a hyperpigmented spot on a skin surface, wherein the composition is applied for a period of time sufficient to improve the appearance of the hyperpigmented spot. The method may include the step of identifying a hyperpigmented spot on a facial skin surface. Other methods as disclosed include a method for improving the appearance of post-inflammatory hyperpigmentation.

FIELD OF THE INVENTION

The present invention relates to compositions comprising banyan tree,lotus, and clover serum fractions. The present invention also relates tomethods for improving the appearance of hyperpigmented mammalian skinusing a synergistic composition comprising banyan tree, lotus, andclover serum fractions.

BACKGROUND OF THE INVENTION

Human skin comprises three principal layers: the epidermis, the dermis,and the subcutaneous fat layer. The epidermis comprises four layers(from top to bottom): the stratum corneum, the granular layer, the spinylayer, and the basal layer. A separate fifth layer, the stratum lucidum,may be present between the stratum corneum and granular layer. The basallayer produces cells which gradually migrate upward to form the otherepidermal layers. As these cells migrate upward, they lose their centralnucleus and start to produce skin proteins (keratins) and fats (lipids).These cells are identified as keratinocytes when present in the upperlayers of the epidermis. Melanocytes are another class of cells locatedin the basal layer of the epidermis. Melanocytes are responsible for theproduction of melanin, which is primary factor in skin pigmentation.

Melanin is produced by a complex set of reactions within the melanocyteinvolving, at a basic level, the enzyme tyrosinase and L-tyrosine as asubstrate. Tyrosinase catalyzes the conversion of L-tyrosine to DOPA(L-3,4-dihydroxyphenylalanine) and of DOPA to dopaquinone. Dopaquinoneundergoes further conversion to form melanin. Melanin aggregates inorganelles known as the melanosomes which are transferred tokeratinocytes along slender filaments of the melanocyte known asdendrites. There are approximately 1500 gene products expressed inmelanosomes with 600 of them being expressed at any given time and 100of them believed to be unique to the melanosome. In addition, there aremany regulatory elements involved in signaling, in the transport ofmelanosomes within the melanocyte, and in the transfer of melanosomes tothe keratinocytes.

The production of melanin can be triggered by a variety of external andinternal events. For example, melanocytes produce additional melaninwhen skin is subjected to UV radiation. The melanin is then transportedvia melanasomes to the keratinocytes, which then leaves the skin with a“tanned” appearance. Once the UV light is removed the melanocytes returnto normal levels of melanin production. Inflammation may initiatehyperpigmentation by direct stimulation of the melanocytes by mediatorssuch as IL-1, endothelin-1, and/or stem cell factor. Reactive oxygenspecies, such as superoxide and nitric oxide, generated in damaged skinor released as by-products from inflammatory cells may be stimulators ofmelanocytes.

Over time, chronic UV exposure and other intrinsic and extrinsic agingfactors may lead to permanent gene expression changes in keratinocytesand/or melanocytes resulting in age-related hyperpigmented spots. ThemRNA levels of some melanogenesis associated genes (for example,tyrosinase, TYRP1) are reported to be increase actinic lentigos (agespots). There may also be accentuation of the epidermal endothelincascade and a role for stem cell factor in hyperpigmentation. Thesechanges can result in overproduction of melanin and resultanthyperpgimented spots that persist even when an insult, such as UVexposure, is avoided.

Even beyond hyperpigmented spots, chronic UV exposure and otherintrinsic and extrinsic aging factors may lead to more subtle changes inskin tone. Often these changes are described as uneven tone or as amottled appearance.

Age spots and hyperpigmented skin tone can add several years ofperceived age to an individual. Thus, there is a continuing desire toprovide compositions and methods of treatment that can improve theappearance of hyperpigmented spots and overall skin tone.

SUMMARY OF THE INVENTION

The present invention relates to compositions comprising banyan tree,lotus, and clover serum fractions and mixtures and/or combinationsthereof. In one embodiment, the composition of the invention comprisesbanyan tree, lotus and clover serum fractions.

A method of improving the appearance of hyperpigmented skin tonecomprising the step of applying a first composition comprising aneffective amount of banyan tree serum fraction, lotus serum fraction,and clover serum fraction blend to an area of hyperpigmented skin,wherein the composition is applied for a period of time sufficient toimprove the appearance of the hyperpigmented skin.

A method of improving the appearance of a hyperpigmented spot comprisingthe step of applying a first composition comprising an effective amountof banyan tree serum fraction, lotus serum fraction, and clover serumfraction blend to a hyperpigmented spot on a skin surface, wherein thecomposition is applied for a period of time sufficient to improve theappearance of the hyperpigmented spot.

A method of improving the appearance of a hyperpigmented spot comprisingthe steps of (a) identifying a hyperpigmented spot on a facial skinsurface and (b) applying a composition comprising an effective amount ofbanyan tree serum fraction, lotus serum fraction, and clover serumfraction blend to the hyperpigmented spot on the facial skin surface,wherein the composition is applied for a period of time sufficient toimprove the appearance of the hyperpigmented spot.

A method of improving the appearance of post-inflammatoryhyperpigmentation, the method comprising the steps of (a) identifying anarea of post-inflammatory hyperpigmentation on a skin surface; (b)applying to the area an effective amount of banyan tree serum fraction,lotus serum fraction, and clover serum fraction blend, wherein thecomposition is applied at least daily for a period of time sufficient toimprove the appearance of the area of post-inflammatoryhyperpigmentation; and (c) applying to the area an effective amount ofan anti-inflammatory agent.

In response to the technical problems identified in the background, thepresent invention may take other forms. Further forms of the presentinvention will be appreciated in the detailed description that follows.

DETAILED DESCRIPTION OF THE INVENTION

All percentages and ratios used herein are by weight of the totalcomposition and all measurements made are at 25° C., unless otherwisedesignated. All numeric ranges are inclusive of narrower ranges;delineated upper and lower range limits are interchangeable to createfurther ranges not explicitly delineated.

The compositions of the present invention can comprise, consistessentially of, or consist of, the essential components as well asoptional ingredients described herein. As used herein, “consistingessentially of means that the composition or component may includeadditional ingredients, but only if the additional ingredients do notmaterially alter the basic and novel characteristics of the claimedcompositions or methods.

The term “apply” or “application” as used in reference to a composition,means to apply or spread the compositions of the present invention ontoa human skin surface such as the epidermis.

The term “dermatologically acceptable” as used herein means that thecompositions or components described are suitable for use in contactwith human skin tissue without undue toxicity, incompatibility,instability, allergic response, and the like.

The term “effective amount” as used herein means an amount of a compoundor composition sufficient to significantly induce a positive benefit.

The term “post-inflammatory hyperpigmentation” as used herein refers toan acute to chronic increase in pigmentation as a response to atransient inflammatory event. Post-inflammatory hyperpigmentation isparticularly prevalent in, but not limited to, dark skin subjects.Post-inflammatory hyperpigmentation typically subsides once thetransient inflammatory event dissipates. Examples of transientinflammatory events include, but are not limited to, acne lesions,ingrown hairs, scratches, insect bites, surfactant damage, andshort-term UV exposure.

The term “hyperpigmented spot” as used herein refers to a defined areaof skin wherein the pigmentation is greater than that of an adjacentarea of skin due to localized and chronic or systemic overproduction ofmelanin. Hyperpigmented spots typically are between about 2 mm and about10 mm in diameter but smaller or larger spots are possible.Hyperpigmented spots can include one or more of age spots, sun spots,solar lentigos, hypo-melanotic lesions, freckles, and melasma spots.

The term “age spots” as used herein refers to a hyperpigmented spotwherein the pigmentation is due to localized and chronic overproductionof melanin caused by intrinsic or extrinsic aging factors.

The term “skin tone agent” as used herein refers to an agent thatregulates melanin production signals, synthesis of melanin, systemictransfer of melanin between the melanocyte and the keratinocyte, and/ormelanin degradation. Skin tone agents can improve the appearance ofuneven skin tone by acting as a lightening or pigmentation reductioncosmetic agent.

The term “skin tone” as used herein refers to the overall appearance ofmelanin in the skin caused by the systemic, rather than transient,synthesis of melanin. Skin tone is typically characterized over a largerarea of the skin. The area ideally may be than 100 mm², but larger areasare envisioned such as the entirety of the facial skin or any of thefacial skin surfaces.

Skin tone can be measured by image analysis. For example, overalllightness can be measured by L* coordinate in L*a*b* color space(International Commission on Illumination). Chromophore mapping such asmelanin mapping and melanin concentration may be used as an indicator ofoverall skin tone. Mean melanin may be calculated from the chromophoremap data. Additionally, skin tone evenness can be determined by melaninevenness which also may be calculated from the chromophore map data.Suitable chromophore mapping techniques are discussed in the examplebelow.

The term “facial skin surface” as used herein refers to one or more offorehead, periorbital, cheek, perioral, chin, and nose skin surfaces.

As used herein, “exogenous solvent” means any solvent that is notinherently present in the plant material, but is placed in contact withthe plant material for the purpose of separating (e.g., extracting)compounds from the plant material.

I. Compositions

The present invention relates to various compositions and, morespecifically, to compositions for application to a skin surface. Thecompositions may be in a wide variety of product forms that include, butare not limited to, solutions, suspensions, lotions, creams, gels,toners, sticks, pencil, sprays, aerosols, ointments, cleansing liquidwashes and solid bars, shampoos and hair conditioners, pastes, foams,powders, mousses, shaving creams, wipes, strips, patches,electrically-powered patches, wound dressing and adhesive bandages,hydrogels, film-forming products, facial and skin masks (with andwithout insoluble sheet), make-up such as foundations, eye liners, andeye shadows, and the like. The composition form may follow from theparticular dermatologically acceptable carrier chosen, if present in thecomposition.

A. Serum Fractions

Compositions of the present invention comprise effective amounts ofbanyan tree serum fraction, lotus serum fraction, and clover serumfraction. As demonstrated by Example 2 below, Applicant has surprisinglyfound that the combination of these three serum fractions results in asynergistic mixture that inhibits the synthesis of melanin, therebyimproving the appearance of skin hyperpigmentation, to a much greaterdegree than what would be expected from the additive effects of eachcomponent separately.

In Example 2a, the % melanin synthesis inhibition of the individualbanyan tree, clover, and lotus serum fractions, respectively, wasmeasured as 42%, 16%, and 1%. Thus, one would expect that the additiveeffect of all three serum fractions in a blend would result in 59%melanin synthesis inhibition (42%+16%+1%=59%). However, the actual serumfraction blend surprisingly resulted in 100% melanin synthesisinhibition, which was 41% greater than expected (100%−59%=41% more thanexpected).

In Example 2b, the melanin synthesis inhibition assay was repeated atserum fraction concentration levels that were one-half those in Example2a. Again, the serum fraction combination resulted in a much greater %melanin synthesis inhibition than what one would have expected from thesum of the individual serum fraction results. Here, the % melaninsynthesis inhibition of the individual banyan tree, clover, and lotusserum fractions, respectively, was measured as 0%, 14%, and 7%. The sumof these three results is 21% (0%+14%+7%=21%). The actual serum fractionblend resulted in 63% melanin synthesis inhibition, which was 42%greater than expected from the sum (63%−21%=42%).

Banyan tree, lotus, and clover serum fractions consist essentially ofthe flower, leaf, and stem serum fractions obtained from FicusBenghalensis, Nelumbo Nucifera, and Trifolium Pratense, respectively.The preferred serum fractions are produced by Akzo Nobel SurfaceChemistry LLC of Ossining, N.Y., USA, under the trade names FicusBengalensis Enriched Serum Fraction™ (INCI Name: Ficus IndicaFlower/Leaf/Stem Juice), Lotus Enriched Serum Fraction™ (INCI Name:Nelumbo Nucifera Flower/Leaf/Stem Juice), and Red Clover Enriched SerumFraction™ (INCI Name: Trifolium Pratense (Clover) Flower/Leaf/StemJuice).

In some embodiments, the composition may comprise banyan tree serumfraction in an amount of from 0.001% to 15%, alternatively from 0.002%to 10%, alternately from 0.025% to 10%, in other embodiments from 0.05%to 10%, in others from 0.05% to 5%, and in others from 0.1% to 5%, byweight of the total composition. The composition may comprise lotusserum fraction in an amount of from 0.001% to 15%, alternatively from0.002% to 10%, alternatively from 0.01% to 15%, alternately from 0.025%to 10%, in other embodiments from 0.05% to 10%, in others from 0.05% to5%, and in others from 0.1% to 5%, by weight of the total composition.The composition may comprise clover serum fraction in an amount of from0.001% to 15%, alternatively from 0.002% to 10%, alternatively from0.01% to 15%, alternately from 0.025% to 10%, in other embodiments from0.05% to 10%, in others from 0.05% to 5%, and in others from 0.1% to 5%,by weight of the total composition.

The method for making a serum fraction comprises the steps of: (a)separating cell juice from clean, fresh, un-wilted plant matter toobtain fresh cell juice, wherein no exogenous liquid is added prior orduring said separating; (b) filtering said fresh cell juice to obtainfiber-free cell juice; and (c) fractionating said fiber-free cell juiceto obtain the serum fraction for use herein. Suitable serum fractionpreparation methods are set forth in U.S. Pat. No. 7,442,391, “BioactiveBotanical Cosmetic Compositions and Processes for their Production,” toKoganov, and in co-pending U.S. Provisional Application Ser. No.61/381,748, filed 10 Sep. 2010 by Swanson et al., the disclosures ofwhich are hereby incorporated by reference herein.

The resulting serum fractions have superior bioactivity versustraditionally prepared plant extracts. Unlike traditional extracts, theserum fraction is prepared from fresh plant cell juice that has beenmechanically separated from the rest of the fresh plant material.Importantly, no exogenous solvent (e.g., water, hexane, acetone,ethanol) is added during the juice separation process. The resultingcell juice contains the full spectrum of compounds found in fresh plantmatter, thus the resulting serum fractions contain a much broader rangeof active compounds than do traditional plant extracts, which containonly the narrow range of compounds that can be separated with aparticular solvent.

Furthermore, using fresh plants maintains the integrity of the bioactivecomponents inherently present in the fresh plant matter. Traditionalplant extracts are not prepared from fresh plant matter, but rather fromdried plant material, which has undergone degradation due todehydration. During dehydration, the cell walls are compromised, causingthe degredation of compounds through mechanisms such as hydrolysis,oxidation, polymerization, Maillard reactions, and isomerization. Whenthe dried leaves are extracted, the resulting extract thus containsthese degradation products that were not originally present in the freshplant matter. Accordingly, the composition of the resulting dry leafextract greatly differs from that of fresh juice and the resulting serumfraction.

B. Skin Tone Agent

In some embodiments, it may be desirable to include a skin tone agent inthe composition. The skin tone agents can be included to further improveoverall skin tone. When present, the compositions of the presentinvention contain up to about 50%, 40%, 30%, 20%, 10%, 5%, or 3%, byweight of the composition, of the skin tone agent. When present, thecompositions of the present invention contain at least about 0.001%,0.01%, 0.1%, 0.2%, 0.5%, or 1%, by weight of the composition, of theskin tone agent. Suitable ranges include any combination of the lowerand upper limits including suitable ranges from about 0.1% to about 50%;from about 0.2% to about 20%; or from about 1% to about 10%, by weightof the composition, of the skin tone agent. The amounts listed hereinare only to be used as a guide, as the optimum amount of the skin toneagent will depend on the specific active selected since their potencydoes vary considerably.

Suitable skin tone agents include, but are not limited to, sugar amines,vitamin B3 compounds, arbutin, deoxyarbutin,1,3-dihydroxy-4-alkylbenzene such as hexylresorcinol, sucrosedilaurante, bakuchoil(4-[(1E, 3S)-3-ethenyl-3,7-dimethyl-1,6octadienyl]phenol or monterpene phenol), pyrenoine (available fromBiotech Marine, France), panicum miliaceum seed extract, arlatone dioicacid, cinnamic acid, ferulic acid, achromaxyl, methyl nicotinamide, oilsoluble licorice extract, folic acid, undecylenic acid (i.e., undecenoicacid), zinc undecylenate, thiamine (Vitamin B1) and its hydrochloride,L-tryptophan, helianthus annuus (sunflower) and vitis vinifera (grape)leaf extract, carnosine (i.e., dragosine), methyl gentisate,1,2-hexandiol and 1,2-octandiol (i.e., combination sold as Symdiol 68 bySymrise AG, Germany), inositol, decylenoylphenylalanine (e.g., soldunder the tradename Sepiwhite by Seppic, France), koijic acid,hexamidine compounds, salicylic acid, and retinoids including retinoland retinol propionate.

In certain embodiments, the additional skin tone agent is selected fromvitamin B3 compounds, sugar amines, hexamidine compounds, salicylicacid, 1,3-dihydroxy-4-alkylbenzene such as hexylresorcinol, andretinoids. As used herein, “vitamin B₃ compound” means a compound havingthe formula:

wherein R is —CONH₂ (i.e., niacinamide), —COOH (i.e., nicotinic acid) or—CH₂OH (i.e., nicotinyl alcohol); derivatives thereof; and salts of anyof the foregoing. As used herein, “sugar amine” includes isomers andtautomers of such and its salts (e.g., HCl salt) and its derivatives.Examples of sugar amines include glucosamine, N-acetyl glucosamine,mannosamine, N-acetyl mannosamine, galactosamine, N-acetylgalactosamine, their isomers (e.g., stereoisomers), and their salts(e.g., HCl salt). As used herein, “hexaminide compound” means a compoundhaving the formula:

wherein R¹ and R² are optional or are organic acids (e.g., sulfonicacids, etc.). In one embodiment, hexamidine compound is hexamidinediisethionate.

C. Anti-Inflammatory Agents

Hyperpigmentation may result from skin inflammation. Transientinflammatory events triggering hyperpigmentation and, more specifically,post-inflammatory hyperpigmentation include, but are not limited to,acne lesions, ingrown hairs, scratches, insect bites, surfactant damage,allergens, and short-term UV exposure. Inflammation inducedhyperpigmentation including post-inflammatory hyperpigmentation may bemanaged by incorporating into the compositions of the present inventionan anti-inflammatory agent. When present, the compositions of thepresent invention contain up to about 20%, 10%, 5%, 3%, or 1% by weightof the composition, of the anti-inflammatory agent. When present, thecompositions of the present invention contain at least about 0.001%,0.01%, 0.1%, 0.2%, 0.3%, 0.5%, or 1%, by weight of the composition, ofthe anti-inflammatory agent. Suitable ranges include any combination ofthe lower and upper limits. Suitable anti-inflammatory agents include,but are not limited to nonsteroidal anti-inflammatory agents (NSAIDSincluding but not limited to ibuprofen, naproxen, flufenamic acid,etofenamate, aspirin, mefenamic acid, meclofenamic acid, piroxicam andfelbinac), glycyrrhizic acid (also known as glycyrrhizin, glycyrrhixinicacid, and glycyrrhetinic acid glycoside) and salts such as dipotassiumglycyrrhizate, glycyrrhetenic acid, licorice extracts, bisabolol (e.g.,alpha bisabolol), manjistha (extracted from plants in the genus Rubia,particularly Rubia cordifolia), and guggal (extracted from plants in thegenus Commiphora, particularly Commiphora mukul), kola extract,chamomile, clover extract (e.g., red clover extract), and sea whipextract (extracts from plant in the order Gorgonacea), derivatives ofany of the foregoing, and mixtures thereof.

D. Sunscreen Actives

The compositions of the subject invention may comprise one or moresunscreen actives (or sunscreen agents) and/or ultraviolet lightabsorbers. Herein, “sunscreen active” collectively includes, sunscreenactives, sunscreen agents, and/or ultraviolet light absorbers. Sunscreenactives include both sunscreen agents and physical sunblocks. Sunscreenactives may be organic or inorganic. Examples of suitable sunscreenactives are disclosed in Personal Care Product Council's InternationalCosmetic Ingredient Dictionary and Handbook, Thirteenth Edition, as“sunscreen agents.” Particularly suitable sunscreen actives are2-ethylhexyl-p-methoxycinnamate (commercially available as PARSOL™ MCX),4,4′-t-butyl methoxydibenzoyl-methane (commercially available as PARSOL™1789), 2-hydroxy-4-methoxybenzophenone, octyldimethyl-p-aminobenzoicacid, digalloyltrioleate, 2,2-dihydroxy-4-methoxybenzophenone,ethyl-4-(bis(hydroxypropyl))aminobenzoate,2-ethylhexyl-2-cyano-3,3-diphenylacrylate, 2-ethylhexyl-salicylate,glyceryl-p-aminobenzoate, 3,3,5-tri-methylcyclohexylsalicylate, menthylanthranilate, p-dimethyl-aminobenzoic acid or aminobenzoate,2-ethylhexyl-p-dimethyl-amino-benzoate, 2-phenylbenzimidazole-5-sulfonicacid, 2-(p-dimethylaminophenyl)-5-sulfonicbenzoxazoic acid, octocrylene,zinc oxide, benzylidene camphor and derivatives thereof, titaniumdioxide, and mixtures thereof.

In one embodiment, the composition may comprise from about 1% to about20%, and alternatively from about 2% to about 10% by weight of thecomposition, of the sunscreen active. Exact amounts will vary dependingupon the chosen sunscreen active and the desired Sun Protection Factor(SPF), which is within the knowledge of one of skilled in the art.

E. Optional Components

The compositions of the present invention may contain a variety of otheringredients provided that they do not unacceptably alter the benefits ofthe invention. When present, compositions of the present invention maycontain from about 0.0001% to about 50%; from about 0.001% to about 20%;or, alternately, from about 0.01% to about 10%, by weight of thecomposition, of the optional components. The amounts listed herein areonly to be used as a guide, as the optimum amount of the optionalcomponents used in a composition will depend on the specific activeselected since their potency does vary considerably. Hence, the amountof some optional components useful in the present invention may beoutside the ranges listed herein.

The optional components, when incorporated into the composition, shouldbe suitable for use in contact with human skin tissue without unduetoxicity, incompatibility, instability, allergic response, and the like.The compositions of the present invention may include optionalcomponents such as anti-acne actives, desquamation actives,anti-cellulite agents, chelating agents, flavonoids, tanning active,non-vitamin antioxidants and radical scavengers, hair growth regulators,anti-wrinkle actives, anti-atrophy actives, minerals, phytosterolsand/or plant hormones, N-acyl amino acid compounds, antimicrobial orantifungal actives, and other useful skin care actives, which aredescribed in further detail in U.S. application publication No.US2006/0275237A1 and US2004/0175347A1, the disclosures of which arehereby incorporated by reference herein.

The Personal Care Product Council's International Cosmetic IngredientDictionary and Handbook, Thirteenth Edition, describes a wide variety ofnon-limiting cosmetic and pharmaceutical ingredients commonly used inthe skin care industry, which are suitable optional components for usein the compositions of the present invention. Examples of theseingredient classes include: abrasives, absorbents, aesthetic componentssuch as fragrances, pigments, colorings/colorants, essential oils,anti-caking agents, antifoaming agents, antimicrobials, binders,biological additives, buffering agents, bulking agents, chelatingagents, chemical additives, colorants, cosmetic astringents, cosmeticbiocides, denaturants, drug astringents, emollients, externalanalgesics, film formers or materials, opacifying agents, pH adjusters,preservatives, propellants, reducing agents, sequestrants, skin coolingagents, skin protectants, thickeners viscosity modifiers, vitamins, andcombinations thereof.

F. Dermatologically Acceptable Carrier

The compositions of the present invention may also comprise adermatologically acceptable carrier (which may be referred to as“carrier”) for the composition. The phrase “dermatologically acceptablecarrier”, as used herein, means that the carrier is suitable for topicalapplication to the keratinous tissue, has good aesthetic properties, iscompatible with the actives in the composition, and will not cause anyunreasonable safety or toxicity concerns. In one embodiment, the carrieris present at a level of from about 50% to about 99%, about 60% to about98%, about 70% to about 98%, or, alternatively, from about 80% to about95%, by weight of the composition.

The carrier can be in a wide variety of forms. Non-limiting examplesinclude simple solutions (e.g., aqueous, organic solvent, or oil based),emulsions, and solid forms (e.g., gels, sticks, flowable solids, oramorphous materials). In certain embodiments, the dermatologicallyacceptable carrier is in the form of an emulsion. Emulsion may begenerally classified as having a continuous aqueous phase (e.g.,oil-in-water and water-in-oil-in-water) or a continuous oil phase (e.g.,water-in-oil and oil-in-water-in-oil). The oil phase of the presentinvention may comprise silicone oils, non-silicone oils such ashydrocarbon oils, esters, ethers, and the like, and mixtures thereof.

The aqueous phase typically comprises water. However, in otherembodiments, the aqueous phase may comprise components other than water,including but not limited to water-soluble moisturizing agents,conditioning agents, anti-microbials, humectants and/or otherwater-soluble skin care actives. In one embodiment, the non-watercomponent of the composition comprises a humectant such as glycerinand/or other polyols. However, it should be recognized that thecomposition may be substantially (i.e., less than 1% water) or fullyanhydrous.

A suitable carrier is selected to yield a desired product form.Furthermore, the solubility or dispersibility of the components (e.g.,extracts, sunscreen active, additional components) may dictate the formand character of the carrier. In one embodiment, an oil-in-water orwater-in-oil emulsion is preferred.

Emulsions may further comprise an emulsifier. The composition maycomprise any suitable percentage of emulsifier to sufficiently emulsifythe carrier. Suitable weight ranges include from about 0.1% to about 10%or about 0.2% to about 5% of an emulsifier, based on the weight of thecomposition. Emulsifiers may be nonionic, anionic or cationic. Suitableemulsifiers are disclosed in, for example, U.S. Pat. No. 3,755,560, U.S.Pat. No. 4,421,769, and McCutcheon's Detergents and Emulsifiers, NorthAmerican Edition, pages 317-324 (1986), the disclosures of which arehereby incorporated by reference herein. Suitable emulsions may have awide range of viscosities, depending on the desired product form.

The carrier may further comprise a thickening agent as are well known inthe art to provide compositions having a suitable viscosity andrheological character.

II. Methods of Treatment

Various methods of treatment, application, regulation, or improvementmay utilize the aforementioned compositions. In one embodiment, themethod includes the step of identifying a hyperpigmented spot forimprovement by the composition. The hyperpigmented spot may beidentified by the user or a third party such as a dermatologist,cosmetician, or other caregiver. Identification may be done by visualinspection of the skin for hyperpigmented spots in need of treatmentbased on size and/or color. Identification may also be done bycommercially available imaging devices such SIAscope® V (available fromAstron Clinica, Ltd., UK) or the VISIA® Complexion Analysis system(available from Canfield Scientific, Inc., Fairfield, N.J.). Bothdevices are capable of collecting images of the skin and identifyinghyperpigmented spots. In some instances, the method comprises the stepof identifying a plurality of hyperpigmented spots for treatment by thecomposition.

Identification of the hyperpigmented spot may occur on any skin surfaceof the body. Skin surfaces of the most concern tend to be those nottypically covered by clothing such as facial skin surfaces, hand and armskin surfaces, foot and leg skin surfaces, and neck and chest skinsurfaces (e.g., décolletage). In particular, identification of thehyperpigmented spot may be on a facial skin surface including theforehead, perioral, chin, periorbital, nose, and/or cheek skin surfaces.

The method may comprise the step of applying the composition to ahyperpigmented spot or spots, which may have been previously identified.Many regimens exist for the application of the composition to thehyperpigmented spot. The composition may be applied at least once a day,twice a day, or on a more frequent daily basis, during a treatmentperiod. When applied twice daily, the first and second applications areseparated by at least 1 to about 12 hours. Typically, the compositionmay be applied in the morning and/or in the evening before bed.

The treatment period is ideally of sufficient time to provide animprovement in the hyperpigmented spot. The improvement may be adetectable reduction in size of the hyperpigmented spot, lightening ofthe hyperpigmented spot (e.g., lighter in color), or decrease in melaninof the hyperpigmented spot. The treatment period may be at least about 1week. The treatment period may last about 4 weeks or about 8 weeks. Incertain embodiments, the treatment period will extend over multiplemonths (i.e., 3-12 months) or multiple years. In one embodiment thecomposition is applied to the hyperpigmented spot(s) at least once a dayduring a treatment period of at least about 4 weeks or at least about 8weeks. In one embodiment the composition is applied to thehyperpigmented spot(s) twice a day during a treatment period of at leastabout 4 weeks or 8 weeks.

The step of applying the composition to the hyperpigmented spot may beaccomplished by localized application. In reference to application ofthe composition, the term “localized”, “local”, or “locally” mean thatthe composition is delivered the targeted area (such as thehyperpigmented spot) while minimizing delivery to skin surface notrequiring treatment. The composition may be applied and lightly massagedinto the hyperpigmented spot. It is recognized that localizedapplication does allow for a reasonable amount of the composition to beapplied to areas adjacent the hyperpigmented spot (i.e., the compositionis unlikely to be applied or to remain within the boundary of thehyperpigmented spot without some spreading). The form of the compositionor the dermatologically acceptable carrier should be selected tofacilitate localized application. While certain embodiments of thepresent invention contemplate applying a composition locally to ahyperpigmented spot, it will be appreciated that compositions of thepresent invention can be applied more generally or broadly to one ormore facial skin surfaces to reduce the appearance of hyperpigmentedspots within those facial skin regions.

In some embodiments, the composition may be delivered by a variety ofapplicators appropriate for localized and general application. Suchapplicators can include droppers, applicator wands, cotton swabs, or anyother suitable device. Other suitable applicators include SH-0127 penapplicator available from Shya Hsin Plastic Works, Inc., Taiwan andeither the Xpress Tip or liquid filled swab available from SwabPlus,Inc., China. The applicator may be configured to easily apply thecomposition to hyperpigmented spots having an approximate diameterbetween about 2 mm and about 10 mm and allowing for a dosed amount ofthe composition of between about 1 to about 50 uL/cm² or between about 1to about 5 uL/cm². In another embodiment, the composition is applied tothe one or more hyperpigmented spots and more generally to one or morefacial skin surfaces contemporaneously (i.e., over a period of less than30 minutes or, more typically, less than 5 minutes).

While some methods described herein contemplate applying thecompositions of the present invention with an applicator, it will beappreciated that applicators are not required and the compositions ofthe present invention can also be applied directly by using one's fingeror in other conventional manners.

In one embodiment, the method comprises the steps of applying a firstcomposition comprising an effective amount of banyan tree serumfraction, lotus serum fraction, and clover serum fraction to ahyperpigmented spot or a plurality of hyperpigmented spots on a skinsurface and of applying a second composition to the skin surface, beforeor after the first composition. The first and second compositions may beany compositions described herein; however, the second composition mayoptionally comprise an effective amount of the serum fractions presentin the first composition. The second composition may comprise one ormore tone agents, sunscreen actives, anti-inflammatory agents, oroptional components. The first composition may be locally applied to thehyperpigmented spot or plurality of hyperpigmented spots. The secondcomposition may be locally applied to the hyperpigmented spot or aplurality of hyperpigmented spots to which the first composition isapplied or the second composition may be applied more generally to theskin surface including the hyperpigmented spots to which the firstcomposition is applied. In certain embodiments, the skin surface isfacial skin surface which include one or more of the forehead, perioral,chin, periorbital, nose, and cheek skin surfaces. In another embodiment,the first and second compositions are applied contemporaneously to atleast the cheek, forehead, and chin/perioral skin surfaces. For generalapplication to a skin surface and, particularly a facial skin surface,the dosed amount of the first or second composition may be between about1 to about 50 uL/cm² per application (i.e., per single application tothe skin surfaces).

Suitable methods may comprise any one or more of the abovementionedsteps. All of the aforementioned steps are applicable to application,treatment, regulation, and/or improvement of both a singlehyperpigmented spot as well as a plurality of hyperpigmented spots.Likewise, the exemplary methods that follow are applicable to both asingle hyperpigmented spot as well as a plurality of hyperpigmentedspots.

One suitable method of improving the appearance of a hyperpigmented spotincludes the step of topically applying a composition comprising aneffective amount of banyan tree serum fraction, lotus serum fraction,and clover serum fraction to the hyperpigmented spot on a skin surface,wherein the composition is applied for a period of time sufficient forthe serum fractions to improve the appearance of the hyperpigmentedspot. Another suitable method of improving the appearance ofhyperpigmented spots includes the steps of identifying a hyperpigmentedspot on a skin surface, applying a composition comprising an effectiveamount of said serum fractions to the hyperpigmented spot on the skinsurface, wherein the composition is applied for a period of timesufficient for the serum fractions to improve the appearance of thehyperpigmented spot.

Another suitable method is for improving the appearance of apost-inflammatory hyperpigmentation. The method may comprise the stepsof identifying an area of post-inflammatory hyperpigmentation on a skinsurface and of applying to the area said serum fractions and ananti-inflammatory active. An effective amount of said serum fractionsmay be applied at least daily for a period of time sufficient to improvethe appearance of the area of post-inflammatory hyperpigmentation. Saidserum fractions may be provided in a first composition and theanti-inflammatory agent provided in a second composition. Alternately,the said serum fractions and the anti-inflammatory agent may be providedin the same composition. The compositions may further comprise asunscreen active, a skin tone agent, or combinations thereof.

EXAMPLES Example 1 Exemplary Compositions

Table 1 sets forth non-limiting examples of the compositions of thepresent invention. The examples are given solely for the purpose ofillustration and are not to be construed as limitations of the presentinvention, as many variations thereof are possible without departingfrom the spirit and scope of the invention, which would be recognized byone of ordinary skill in the art. In the examples, all concentrationsare listed as weight percent, unless otherwise specified and may excludeminor materials such as diluents, filler, and so forth. The listedformulations, therefore, comprise the listed components and any minormaterials associated with such components. As is apparent to one ofordinary skill in the art, the selection of these minor materials willvary depending on the physical and chemical characteristics of theparticular ingredients selected to make the present invention asdescribed herein.

All examples may be used to treat or improve the appearance of one ormore hyperpigmented spots. The present invention may further relate to aregimen involving the localized treatment for one or more hyperpigmentedspots by a first composition (e.g., Examples A or B) and a more broad orgeneral facial skin treatment by a second composition (e.g., Examples Cor D), which can be applied before or after the localized treatment toimprove skin tone across the face.

TABLE 1 Exemplary Compositions Component/% by wt. Example A Example BExample C Example D Ficus Benghalensis 0.55 1.00 0.55 0.00 (Banyan Tree)Flower/Leaf/Stem Juice (**) Nelumbo Nucifera (Lotus) 0.10 0.20 0.20 0.00Flower/Leaf/Stem Juice (**) Trifolium Pratense (Clover) 0.10 0.20 0.200.00 Flower/Leaf/Stem Juice (**) N-Acetylglucosamine 0.00 0.00 2.00 0.00Hexamidine Diisethionate 0.00 0.00 0.09 0.09 Sepiwhite ™ 0.00 0.00 0.500.50 (Undecylenoyl- phenylalanine, neutralized) (available from SEPPIC)Sepigel 305 ™ 0.00 0.00 2.00 2.00 (Polyacrylamide + C13-14 isoparaffin +laureth-7) (available from SEPPIC) Dipotassium Glycyrrhizate 0.00 0.100.10 0.30 Hexamidine Diisethionate 0.00 0.00 0.09 0.09 Homosalate 0.000.00 0.00 9.00 Avobenzone 0.00 0.00 0.00 3.00 Octocrylene 0.00 0.00 0.002.60 Oxybenzone 0.00 0.00 0.00 1.00 Octisalate 0.00 0.00 0.00 4.50Butylene Glycol (CAS No. 5.50 5.50 5.50 5.50 107-88-0) Niacinamide (CASNo. 98- 5.00 5.00 5.00 5.00 92-0) Glycerin (CAS No. 56-81- 2.50 2.502.50 2.50 5) DC 1503 Fluid ™ (available 2.50 2.50 2.50 2.50 fromDowCorning) Lubrajel Oil ™ (available 1.44 1.44 1.44 1.44 from Sederma)Phenonip XB ™ (available 1.25 1.25 1.25 1.25 from Clariant) D-panthenol(CAS No. 81- 1.00 1.00 1.00 1.00 13-0) Tospearl 2000 ™ 1.00 1.00 1.001.00 (Polymethylsils esquioxane) (CAS No. 68554-70-1) (available from GESilicones/Momentive) DL-Alpha Tocopheryl 0.50 0.50 0.50 0.50 Acetate(CAS No. 7695-91- 2) Prodew 400 ™ (available 0.50 0.50 0.50 0.50 fromAjinomoto) Pemulen TR-2 ™ 0.25 0.25 0.25 0.25 (Acrylates/C10-30 AlkylAcrylate Crosspolymer) (available from Noveon) Polysorbate 20 (CAS No.0.25 0.25 0.25 0.25 9005-64-5) Sodium Metabisulfite (CAS 0.25 0.25 0.250.25 No. 7681-57-4) Allantoin (CAS No. 97-59- 0.20 0.20 0.20 0.20 6)Sodium Hydroxide (CAS 0.17 0.17 0.17 0.17 No. 1310-73-2) (50% solutionby weight in water) Disodium EDTA (CAS No. 0.10 0.10 0.10 0.10 139-33-3)Xanthan Gum (CAS No. 0.05 0.05 0.05 0.05 11138-66-2) Sodium Hyaluronate(CAS 0.01 0.01 0.01 0.01 No. 9067-32-7) Water (CAS No. 7732-18- QS QS QSQS 5) TOTAL (% by weight of 100.00 100.00 100.00 100.00 totalcomposition) (**) Manufactured by Akzo Nobel Surface Chemistry LLC ofOssining, New York, USA.

The compositions of the present invention are generally prepared byconventional methods such as are known in the art of making topicalcompositions. Such methods typically involve mixing of the ingredientsin one or more steps to a relatively uniform state, with or withoutheating, cooling, application of vacuum, and the like. Typically,emulsions are prepared by first mixing the aqueous phase materialsseparately from the fatty phase materials and then combining the twophases as appropriate to yield the desired continuous phase. Thecompositions are preferably prepared such as to optimize stability(physical stability, chemical stability, photostability) and/or deliveryof the active materials. This optimization may include appropriate pH(e.g., less than 7), exclusion of materials that can complex with theactive agent and thus negatively impact stability or delivery (e.g.,exclusion of contaminating iron), use of approaches to prevent complexformation (e.g., appropriate dispersing agents or dual compartmentpackaging), use of appropriate photostability approaches (e.g.,incorporation of sunscreen/sunblock, use of opaque packaging), etc.

Example 2 Melanin Synthesis Inhibition

Individual banyan tree, lotus, and clover serum fractions, as well astheir blends, were evaluated according to the melanin synthesis assaydescribed herein. A B16-F1 mouse melanoma cell line is employed in theassay. The B16-F1 cells are obtained from American Tissue CultureCollection, Virginia, USA. The cell culture medium used in the assaycomprises 500 mL of Dulbecco's Modified Eagle's Medium (DMEM), 50 mLFetal Bovine Serum (FBS), and 5 mL of penicillin-streptomycin liquid.B16-F1 cells that are cultured in this medium and grown to greater than90% confluency synthesize melanin. While not intending to be bound byany theory, it is hypothesized that the melanin synthesis is stimulatedby the culture medium and/or stress induced by growth to a highconfluency. The DMEM and FBS can be obtained from American TissueCulture Collection and the penicillin-streptomycin liquid can beobtained from Invitrogen, Inc., California, USA. Equipment used in theassay include a CO₂ incubator, such as a Forma Series Model 3110 byTherma Scientific, Massachusets, USA; a Hemocytometer, such as a BrightLine model by Hauser Scientific, Pennsylvania, USA; and a UV-VisibleSpectrum Plate Reader, such as a SpectraMax250 from Molecular Devices,California, USA. The assay steps include:

-   Day 0—Cell Growth: Warm the cell culture medium to 37° C. and place    29 mL into a T-150 flask. Add approximately 1×10⁶ of B16-F1 passage    1 mouse cells to the T-150 flask and incubate for 3 days at 37° C.,    5% CO₂, 90% relative humidity, until about 80% confluency.-   2. Day 3—Initiate a 96 Well Plate: At day 3, trypsinize the cells    from the T-150 flask and determine the concentration of cells using    the Hemacytometer. Initiate a 96 well plate with 2,500 cells per    well in 100 uL of cell culture medium. Incubate the plate at 37° C.,    5% CO₂, 90% relative humidity for 2 days until at least 20% to 40%    confluent.-   3. Day 5—Remove the cell culture medium from the plate and replace    with fresh culture medium (100 uL per well). Add luL of each test    compound. Multiple dilution ratios may be tested in order to    generate a dose response curve, wherein preferably three wells are    treated with each dilution ratio. Controls comprise wells having the    cell culture medium, B16-F1 cells, and the solvent (control #1);    wells comprising the cell culture medium and the solvent (control    #2); and optionally wells comprising the cell culture medium,    solvent and [test compound] when necessary to control for the [test    compound] background color (control #3).-   4. Day 7—Measure Melanin Production: Cells should have a confluency    greater than about 80%. If not, this data point is not used. Add 100    uL of a 0.75% sodium hydroxide solution to each well. Read the 96    well plate using the UV-Vis Plate Reader at 410 nm to optically    measure the amount of melanin produced between wells that are    treated with [test compound] and control wells that are not. Wells    in which melanin is produced appear brownish in color. Wells in    which little melanin is produced appear clear to light purple in    color. Percentage of melanin synthesis inhibition is calculated by    the following equation:

$\frac{100 - \lbrack {{OD}_{410\mspace{14mu} {Test}\mspace{14mu} {Compound}} - {OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 2}}} \rbrack}{( {{OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 1}} - {OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 2}}} )} \times 100$

Where OD₄₁₀ is the Optical Density at 410 nm as measured by the UV-VisSpectrum Plate Reader.

When Control #3 is used, the formula for percentage melanin synthesisinhibition is:

$\frac{100 - \lbrack {{OD}_{410\mspace{14mu} {Test}\mspace{14mu} {Compound}} - {OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 3}}} \rbrack}{( {{OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 1}} - {OD}_{410\mspace{14mu} {Control}\mspace{14mu} {\# 2}}} )} \times 100$

Using generally the assay outlined above, individual serum fractions andblends were analyzed for melanin synthesis in B16-F1 cells. The resultsobtained at two different test concentrations are set forth below asExample 2a and Example 2b.

Example 2a

T-test vs. Blend Serum % Melanin (<0.05 statistically Fraction %Concentration Inhibition significant) Banyan 0.275 42 0.009291 TreeClover 0.05 16 0.007881 Lotus 0.05 1 0.001603 Blend 0.275 banyan tree +0.05 100 clover + 0.05 lotus

Example 2b

T-test vs. Blend Serum % Melanin (<0.05 statistically Fraction %Concentration Inhibition significant) Banyan 0.1375 0 0.001049 TreeClover 0.025 14 0.004098 Lotus 0.025 7 0.006366 Blend 0.275 banyantree + 0.05 63 clover + 0.05 lotus

Example 3 Method of Treatment

A test subject topically applies composition A of Example 1 to theentire face once a day for 8 weeks. Photographs of the subject's face atweek 0 and at week 8 are compared to show that the subject's overallfacial skin tone improves (less hyperpigmented and more even) andhyperpigmented spots are lighter and smaller in size.

The dimensions and values disclosed herein are not to be understood asbeing strictly limited to the exact numerical values recited. Instead,unless otherwise specified, each such dimension is intended to mean boththe recited value and a functionally equivalent range surrounding thatvalue. For example, a dimension disclosed as “40 mm” is intended to mean“about 40 mm.”

Every document cited herein, including any cross referenced or relatedpatent or application, is hereby incorporated herein by reference in itsentirety unless expressly excluded or otherwise limited. The citation ofany document is not an admission that it is prior art with respect toany invention disclosed or claimed herein or that it alone, or in anycombination with any other reference or references, teaches, suggests ordiscloses any such invention. Further, to the extent that any meaning ordefinition of a term in this document conflicts with any meaning ordefinition of the same term in a document incorporated by reference, themeaning or definition assigned to that term in this document shallgovern.

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

What is claimed is:
 1. A serum fraction blend suitable for use informulating topically applied skin compositions, said blend comprising:a. Banyan tree serum fraction; b. Lotus serum fraction; and c. Cloverserum fraction.
 2. The serum fraction blend of claim 1, wherein each ofsaid banyan tree, lotus, and clover serum fractions are produced by theprocess comprising: a. separating plant cell juice from clean, fresh,un-wilted plant matter to obtain fresh plant cell juice, wherein noexogenous liquid is added prior or during said separating; b. filteringsaid fresh plant cell juice to obtain fiber-free cell juice; and c.fractionating said fiber-free cell juice to obtain said serum fractions.3. The serum fraction blend of claim 1 which comprises: a. from 0.001%to 15% Banyan tree serum fraction; b. from 0.001% to 15% Lotus serumfraction; and c. from 0.001% to 15% Clover serum fraction.
 4. The serumfraction blend of claim 1 which comprises: a. from 0.002% to 10% Banyantree serum fraction; b. from 0.002% to 10% Lotus serum fraction; and c.from 0.002% to 10% Clover serum fraction.
 5. The serum fraction blend ofclaim 1 which comprises: a. from 0.025% to 10% Banyan tree serumfraction; b. from 0.025% to 10% Lotus serum fraction; and c. from 0.025%to 10% Clover serum fraction.
 6. The serum fraction blend of claim 1which comprises: a. from 0.05% to 5% Banyan tree serum fraction; b. from0.05% to 5% Lotus serum fraction; and c. from 0.05% to 5% Clover serumfraction.
 7. The serum fraction blend of claim 1 wherein said blend iseffective in inhibiting melanin synthesis, wherein the percentage ofmelanin synthesis inhibition for said blend is greater than thepercentage of melanin synthesis inhibition for each of the individualblend components when added together.